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© COPYRIG
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UNIVERSITI PUTRA MALAYSIA
EXTRACTION OF STARCH, XYLOSE AND GLUCLOSE FROM OIL PALM STEM USING CHEMICAL STEEPING AND DILUTE ACID HYDROLYSIS
METHODS
WONG LIH JIUN
FH 2011 14
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EXTRACTION OF STARCH, XYLOSE AND
GLUCOSE FROM OIL PALM STEM USING
CHEMICAL STEEPING AND DILUTE ACID
HYDROLYSIS METHODS
WONG LIH JIUN
MASTER OF SCIENCE
UNIVERSITI PUTRA MALAYSIA
2011
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EXTRACTION OF STARCH, XYLOSE AND GLUCLOSE FROM OIL PALM
STEM USING CHEMICAL STEEPING AND DILUTE ACID HYDROLYSIS
METHODS
By
WONG LIH JIUN
Thesis Submitted to the School of Graduate Studies, Universiti Putra Malaysia,
in Fulfillment of the Requirements for the Degree of Master of Science
July 2011
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DEDICATION
To my beloved parents,
“Wong Choo Koh and Lim Yoke Sim”
sisters,
“Wong Chuan Kuan and Wong Wen Jun”
brother,
“Wong Chao Ching”
And
All my beloved friends…
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Abstract of thesis presented to the Senate of Universiti Putra Malaysia in fulfillment of
the requirement for the degree of Master of Science
EXTRACTION OF STARCH, XYLOSE AND GLUCOSE FROM OIL PALM
STEM USING CHEMICAL STEEPING METHOD AND DILUTE ACID
HYDROLYSIS METHODS
By
WONG LIH JIUN
JULY 2011
Chairman: H’ng Paik San, PhD
Faculty: Faculty of Forestry
In Malaysia, oil palm industry is the largest biomass producers compared to other types
of biomass generated. With the growth of palm oil production in Malaysia, the amount
of oil palm stem generated also shows a corresponding increase. The utilization of oil
palm stem in plywood production is not so economically sound due to great variations in
physical and mechanical properties. As a result, numerous researches and studies on the
chemical derivatives of oil palm trunk have been conducted.
This study focused on the total extractable amount of starch, xylose and glucose from oil
palm stem using chemical steeping method and dilutes acid hydrolysis. The specific
objectives were to determine the chemical composition and the effect of chemical
steeping variables (steeping temperature and steeping hours) on the starch yield, and
acid hydrolysis parameters (particle sizes, acid concentration and reaction time) on
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glucose and xylose yield. This research also served to investigate the effectiveness of
chemical steeping as pre-treatment for dilute acid hydrolysis.
Oil Palm stem were used in powder form of 40 mesh size and dried to 5% moisture
content for chemical composition tests prior to chemical analysis whilst 20 mesh, 40
mesh and 60 mesh sizes were used for chemical steeping and dilute acid hydrolysis.
Chemical analysis was based on TAPPI standard T 203 for determination of its main
composition. Low lignin, hemicelluloses and cellulose content were found in this batch
of study. Hemicellulose content is the one for the most important factors for xylose
production and glucose produced from cellulose from lignocellulosic.
Starch in the oil palm stem was extracted by using chemical steeping method. The oil
palm stem powder (20, 40 or 60 mesh) was steeped with 0.2% of sodium metabisulphite
(Na2S2O5) and 0.5% of lactic acid (C3H6O3) at room temperature (±26˚C), 40˚C and
50˚C for 36, 48 hours respectively. The result showed that higher starch yield was
obtained when steeped with temperature. The optimum starch yield can be obtained by
chemical steeping method under 50˚C for 48 hours.
Two treatments were employed for dilute acid hydrolysis in producing xylose and
glucose from oil palm stem which were carried out 1) without pre-treatment 2) with pre-
treatment. The chemical steeping method was treated as a pre-treatment, which the
optimum variable from steeping was chosen as the pre-treatment variable (50˚C and 48
hours) for dilute acid hydrolysis. Each of the treatments was treated with three different
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sulfuric acid concentrations (2%, 4% and 6%) at two different reaction time (30 minutes
and 60 minutes) at temperature 115˚C.
Analysis of variance revealed statistically significant difference in dilute acid hydrolysis
variables for xylose and glucose yield. Significant interaction among the variables was
observed for glucose yield, whereas no significant interaction for the dilute acid
hydrolysis variable was observed for xylose. For xylose, individual variables were
affecting the xylose yield independently. The Tukey test showed the highest glucose
yield, which is 25.5% (dry w/w) was obtained using the dilute acid hydrolysis with
parameter of 2% sulfuric acid concentration reacted for 30 minutes on 60 meshes
without pre-treated oil palm stem powder. On the other hand, the presence of pre-
treatment before dilute acid hydrolysis process contributed to higher xylose yield
(35.0%) from oil palm stem. The particle sizes of oil palm stem powder (20 mesh, 40
mesh dan saiz 60 mesh) and acid concentration (2%, 4% dan 6%) were not significant
different for xylose yield but higher xylose yield can be extracted with the presence of
pre-treament and hydrolysis for 60 minutes.
Conclusively, the oil palm stem has substantial amount of starch, xylose and glucose.
Starch was effectively extracted from oil palm stem using chemical steeping method.
The presence of chemical steeping as pre-treatment was the most independent variable
in affecting the optimum glucose and xylose yield extracted from oil palm stem.
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Abstrak tesis yang dikemukakan kepada Senat Universiti Putra Malaysia sebagai
memenuhi keperluan Ijazah Master Sains
PENGEKSTRASI KANJI, XYLOSA DAN GLUKOSA DARIPADA BATANG
KELAPA SAWIT DENGAN MENGGUNAKAN KIMIA SEDUHAN DAN
HIDROLISIS ASID CAIR
Oleh
WONG LIH JIUN
JULAI 2011
Pengerusi: H’ng Paik San, PhD
Fakulti: Fakulti Perhutanan
Di Malaysia, industri minyak sawit adalah penghasil biojisim lignoselulosa terbesar jika
dibandingkan dengan biojisim-biojisim yang lain. Dengan pertumbuhan pengeluaran
kelapa sawit di Malaysia, secara tidak langsung jumlah batang kelapa sawit yang
dihasilkan juga menunjukkan peningkatkan. Walaubagaimanapun, pengeluaran batang
kelapa sawit sebagai papan lapis tidak begitu cerah dari segi ekonomi kerana variasi
yang besar dalam sifat fizikal dan mekanikal. Oleh sebab itu, banyak kajian dan
penyelidikan telah dilakukan atas cara-cara penerbitan bahan kimia dari batang kelapa
sawit.
Penyelidikan ini fokus pada jumlah kandungan kanji, xylosa dan glucosa yang dapat
diekstrak dari batang kelapa sawit dengan menggunakan kaedah seduhan bahan kimia
dan hidrolisis asid cair. Objektif spefisik adalah untuk mengenalpasti komposisi bahan
kimia dan mengujikaji pengaruh size serbuk dan parameter pemprosesan (suhu seduhan
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dan masa) ke atas hasil kanji yang diekstrak dari batang kelapa sawit. Untuk hidrolisis
acid cair, kesan saiz serbuk batang kelapa sawit dan parameter hidrolisis (kepekatan acid
sulfurik dan masa hidrolisis) ke atas hasil jumlah xylosa dan glukosa yang diekstrak dari
batang kelapa sawit dinilai. Keberkesanan seduhan bahan kimia sebagai pra-rawatan
untuk hidrolisis asid cair juga dinilai.
Untuk analisis komposisi kimia, batang kelapa sawit dalam bentuk serbuk yang bersaiz
40 mesh dikeringkan sehingga 5% kandungan air bagi tujuan siri analisis kimia
dijalankan berdasarkan piawai Tappi T 203 untuk mendapatkan kandungan lignin,
selulosa dan hemiselulosa dalam batang kelapa sawit. Kandungan lignin yang rendah
dan kandungan hemicellulosa, selulosa didapati pada kajian tersebut. Hemiselulosa
bertanggungjawab untuk pengeluaran xylosa dan selulosa adalah bertanggungjawab
untuk pengeluaran glukosa.
Kandungan kanji diekstrak dari batang kelapa sawit dengan menggunakan kaedah
seduhan bahan kimia. Serbuk batang kelapa sawit (20, 40 atau 60 mesh) direndam
dengan kepekatan 0.2% metabisulfit natrium (Na2S2O5) dan 0.5% laktat asid (C3H6O3)
pada suhu bilik (± 26 ˚ C), 40 ˚ C dan 50 ˚ C; masing-masing selama 36 jam dan 48 jam.
Keputusan menunjukkan bahwa kandungan kanji yang tinggi didapati jika dengan
kehadiran suhu. Dengan itu, hasil kanji yang optimum boleh diperolehi melalui cara
seduhan bahan kimia di bawah keadaan suhu 50˚C selama 48jam.
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Untuk hidrolisis asid cair, dua teknik yang berbeza digunakan; 1) teknik dengan tanpa
pra-rawatan 2) teknik pra-rawatan. Kaedah seduhan kimia yang dianggap sebagai pra-
rawatan diaplikasikan sebelum hidrolisis. Sebelum hidrolisis asid cair, serbuk batang
kelapa sawit dirawat dengan seduhan bahan kimia pada suhu 50˚C selama 48 jam. Setiap
sampel direaksikan dengan tiga jenis kepekatan asid sulfurik yang berbeza (2%, 4% dan
6%) pada dua tempoh reaksi yang berbeza (30 minit dan 60 minit) dengan suhu 115 ˚ C.
Analisis varian menunjukkan bahawa perbezaan signifikasi didapati pada parameter
hidrolisis asid cair dalam penghasilan xylosa dan glukosa. Interaksi signifikasi antara
pelbagai pembolehubah didapati bagi penghasilan glukosa, sedangkan tidak ada
interaksi yang signifikasi bagi penghasilan xylosa. Setiap pembolehubah hidrolisis asid
cair memberi kesan individual ke atas hasil xylosa. Keputusan ujian analisis Tukey
menunjukkan bahawa hasil glukosa yang tertinggi 25.5% (jisim kering/ jisim) diperolehi
daripada 60 mesh serbuk batang kelapa sawit tanpa pra-rawatan dengan parameter
hidrolisis yang menggunakan kepekatan asid sulfurik 2% bereaksi selama 30 minit. Ini
adalah dipercayakan bahawa kandungan glukosa adalah dikonversi daripada kanji dan
ikatan karbon adalah mudah dihidrolisiskan oleh kepekatan asid yang rendah dalam
masa reaksi yang singkat. Umumnya, kewujudan pra-rawatan sebelum proses hidrolisis
asid cair menyumbang ke atas hasil xylosa yang lebih tinggi daripada batang kelapa
sawit. Saiz serbuk batang kelapa sawit (20 mesh, 40 mesh dan saiz 60 mesh) dan
kepekatan asid sulfurik (2%, 4% dan 6%) tidak membawa signifikan yang berbeza untuk
menghasilkan xylosa sedangkan waktu reaksi hidrolisis (30 minit dan 60 minit)
membawa perbezaan yang signifikan bagi hasil xylosa. Keputusan kajian tersebut
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menunjukkan bahawa hasil xylosa yang lebih tinggi boleh dihasilkan dengan pra-
rawatan dan dihidrolisis dalam masa reaksi yang lebih lama iaitu selama 60 minit.
Secara kesimpulannya, batang kelapa sawit mengandungi kandungan kanji, xylosa dan
glucosa yang tinggi. Kandungan kanji dapat diekstrak daripada batang kelapa sawit
dengan menggunakan kaedan seduhan kimia. Selain itu, kaedah seduhan kimia
merupakan pembolehubah yang paling membawa kesan individu untuk mempengaruhi
pengekstrakan hasil optimum glucosa dan xylosa daripada batang kelapa sawit.
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ACKNOWLEDGEMENTS
I would like to take this opportunity to express my sincere thanks and appreciation to the
following persons, who have directly or indirectly given generous contribution toward
the completion of this final project.
First, I would like to dedicate my utmost appreciation to my supervisor, Dr. H’ng Paik
San for his encouragement, guidance, invaluable help, and advices throughout the period
of this study. I am also very grateful indeed to my another supervisor, Assoc. Prof. Dr.
Tey Beng Ti for his kind assistance, encouragement and insightful comments.
My appreciations also extend to my especially best of friends and my course mates Miss
Chin Kit Ling, Miss Tey Pei Wen and all who have encouraged and guided me.
Last but not least, my deepest appreciation goes to my beloved parents, sisters, brother
for their undying love, concern and moral support.
Thank You.
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I certify that a Thesis Examination Committee has met on 26th
July 2011 to conduct the
final examination of Wong Lih Jiun on her thesis entitled “Extraction of Starch,
Xylose and Glucose from Oil Palm Stem Using Chemical Steeping and Dilute Acid
Hydrolysis Methods.” In accordance with the Universities and University Colleges Act
1971 and the Constitution of the University Putra Malaysia [P.U.(A) 106] 15 March
1998. The Committee recommends that the student be awarded the Master of Science.
Member of the Examination committee were as follows:
Mohd Zin bin Jusoh
Associate Professor
Faculty of Forestry
Universiti Putra Malaysia
(Chairman)
Dr Hamami bin Sahri, PhD
Professor
Faculty of Forestry
Universiti Putra Malaysia
(Internal Examiner)
Luqman Chuah Bin Abdullah, PhD
Professor
Faculty of Engineering
Universiti Putra Malaysia
(Internal Examiner)
Mariusz Lukasz Maminski, Phd
Lecturer
Warsaw University of Life Science
Poland
(External Examiner)
___________________________
SEOW HENG FONG, PhD
Professor and Deputy Dean
School of Graduate Studies
Universiti Putra Malaysia
Date: 20 December 2011
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This thesis was submitted to the Senate of Universiti Putra Malaysia and has been
accepted as fulfilment of the requirements for the degree of Master of Science. The
members of the Supervisory Committee were as follows:
H’ng Paik San, PhD
Faculty of Forestry
University Putra Malaysia
(Chairman)
Tey Beng Ti, PhD
Associate Professor
Faculty of Engineering
University Putra Malaysia
(Member)
________________________________
BUJANG BIN KIM HUAT, PhD
Professor and Dean
School of Graduate Studies
University Putra Malaysia
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DECLARATION
I declare the thesis is my original work except for quotations and citations which have
been duly acknowledged. I also declare that it has not been previously, and is not
concurrently, submitted for any other degree at University Putra Malaysia or at any other
institution.
_____________________
WONG LIH JIUN
Date: 26 JULY 2011
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TABLE OF CONTENTS
Page
DEDICATION ii
ABSTRACT iii-v
ABSTRAK vi-ix
ACKNOWLEDGEMENTS x
APPROVAL SHEET xi-xii
DECLARATION FORM xiii
LIST OF TABLES xvi
LIST OF FIGURES xvii
LIST OF PLATES xviii
LIST OF ABBREVIATIONS xix
CHAPTER
1 INTRODUCTION
1.1 General Background 1-4
1.2 Statement of Problem 4-7
1.3 Research Objectives 7-8
1.4 Thesis Outline 8
2 LITERATURE REVIEW 2.1 Lignocellulosic Materials 9-10
2.2 Oil Palm Stem as a Lignocellulosic Material 10-11
2.2.1 Availability of Oil Palm Waste in Malaysia 11-12
2.2.2 Utilization of Oil Palm Stem Waste 13-14
2.3 Physical Properties of Oil Palm Stem 14-15
2.4 Chemical Composition of Oil Palm Stem 16
2.4.1 Hemicellulose 17-18
2.5 Starch 18-22
2.4.1 Oil Palm Stem Starch 22-23
2.6 Xylose 24
2.6.1 Xylitol 24-25
2.7 Starch Extraction Method 25
2.7.1 Traditional of Starch Extraction method 26
2.7.2 Steeping Method 26-28
2.8 Hydrolysis 28-29
2.8.1 Hydrolysis Parameter 29-30
2.8.2 Dilute Acid Hydrolysis of Hemicellulose Fibre 30
3 Materials and Methodology
3.1 Materials Preparation 31
3.2 Determination of Chemical Composition 32
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3.3 Chemical Steeping 37
3.3.1 Experimental Design 37
3.3.2 Method Flowchart 38
3.3.3 Method 39
3.3.4 Evaluation 40
3.3.4.1 Standard Reference Curve Calibration 40
3.3.4.2 Starch Determination 41-42
3.4 Dilute Acid Hydrolysis 42
3.4.1 Experimental Design 42-43
3.4.2 Method Flowchart 44
3.4.3 Method 45-48
3.4.4 Analysis Method 48-49
3.4.5 Evaluation 49
4. RESULTS AND DISCUSSION
4.1 General 50
4.2 Chemical Composition of Oil Palm Stem 50-51
4.3 Chemical Steeping 52-55
4.4 Dilute Acid Hydrolysis 56
4.4.1 Glucose Yield 56-59
4.4.2 Xylose Yield 59-62
5 GENERAL CONCLUSION
AND RECOMMENDATIONS
5.1 Conclusions 63-64
5.2 Recommendations 64
REFENRECES 65-72
APPENDIX 1 73
APPENDIX 2 74
BIODATA OF STUDENT 75
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